The subject of the present invention is proteins derived from the coffee bean, and DNAs encoding and regulating the expression of at least one of these proteins.
It is known that numerous plants are capable of producing, in their embryos, in their tubers and in particular in their seeds, storage proteins during their growth. These storage proteins play an important role, in particular, in the storage of amino acids for germination of the seed. They are also important in the structure and the content of amino acids.
Some of these proteins have been isolated and, in some cases, have been expressed in host plants.
Thus, EP 0,295,959 demonstrates, in particular, the expression, in a host plant, of the DNA derived from Bertholletia excelsa H.B.K. (brazil nut) encoding at least one subunit of the storage protein called 2S.
Furthermore, WO 9119801 demonstrates the existence of two storage proteins derived from Theobroma cacao, their precursor and their genes encoding these proteins.
However, up until now, no storage protein derived from the coffee bean and no sequence capable of regulating the transcription of these proteins are known. Yet, it would be very useful to have available sequences of such proteins, in particular in order to modify the original production of the storage proteins in the coffee bean. Furthermore, it would also be very useful to have available a sequence capable of regulating the transcription of such proteins, so as to allow, in particular, the expression, in the coffee bean, of a protein encoded by a gene of interest.
The aim of the present invention is to respond to these needs.
To this effect, the present invention relates to any DNA derived from the coffee bean, encoding at least 20 consecutive amino acids of the amino acid sequence SEQ ID NO:2.
The present invention relates to any storage protein derived from the coffee bean, having at least 20 consecutive amino acids of the amino acid sequence SEQ ID NO:2.
Another subject of the present invention relates to all or part of the DNA delimited by nucleotides 1 to 2509 of the nucleic sequence SEQ ID NO:3, capable of regulating the transcription of the storage proteins according to the invention, as well as the use of all or part of this DNA to direct the expression of genes of interest in plants, in particular in the coffee tree.
The present invention also relates to the use of all or part of the DNA delimited by nucleotides 33 to 1508 of the nucleic sequence SEQ ID NO: 1 or of its complementary strand, of at least 10 bp, to carry out a PCR or as probe to detect in vitro or to inactivate in vivo a coffee bean gene encoding a storage protein.
Furthermore, the invention relates to any recombinant plant cell capable of expressing a recombinant storage protein according to the invention.
Finally, the present invention relates to any food, cosmetic or pharmaceutical product comprising all or part of the DNA or of the recombinant proteins according to the invention.
The present invention therefore opens the possibility of using all or part of the DNA according to the invention so as to modify the original production of the storage proteins in the coffee bean. It is therefore possible in particular to envisage overexpressing or underexpressing the expression of all or part of the DNA according to the invention in the coffee bean.